Li JinBin

Chlorimuronethyl Resistance Selectable Marker Unsuited for the Transformation of Rice Blast Fungus (Magnaporthe Grisea)

Chlorimuronethyl resistance gene is increasingly used as a selectable marker for transformation, especially fungal transformation. Magnaporthe grisea is an important model organism for investigating fungal pathogenicity, and Agrobacterium tumefaciens-mediated transformation (ATMT) is used for functional mutagenesis of the fungus. However, our results showed that rice blast strains collected from infectious rice fields have highly conserved resistance to chlorimuronethyl, even comparable to transformants which carrying chlorimuronethyl resistance genes as selectable marker in laboratory conditions. PCR results showed that all tested field strains presented the amplified products of the same size as the selectable marker amplified from plasmid carrying chlorimuronethyl gene. Sequence analysis of PCR products amplified from field strains confirmed that field strains harbored the highly identity homolog of chlorimuronethyl resistance gene. Blast search in GenBank suggested that the fragment is presenting in reference genome sequence of 70-15, but it is not a wide-spread gene in other organisms, excepted for Herpetosiphon aurantiacus. Although the origin and reason of the conserved chlorimuronethyl resistance gene in field isolates of blast fungus is unclear, the ecological function of the gene is noteworthy.Chlorimuronethyl resistance gene is increasingly used as a selectable marker for transformation, especially fungal transformation. Magnaporthe grisea is an important model organism for investigating fungal pathogenicity, and Agrobacterium tumefaciens-mediated transformation (ATMT) is used for functional mutagenesis of the fungus. However, our results showed that rice blast strains collected from infectious rice fields have highly conserved resistance to chlorimuronethyl, even comparable to transformants which carrying chlorimuronethyl resistance genes as selectable marker in laboratory conditions. PCR results showed that all tested field strains presented the amplified products of the same size as the selectable marker amplified from plasmid carrying chlorimuronethyl gene. Sequence analysis of PCR products amplified from field strains confirmed that field strains harbored the highly identity homolog of chlorimuronethyl resistance gene. Blast search in GenBank suggested that the fragment is presenting in reference genome sequence of 70-15, but it is not a wide-spread gene in other organisms, excepted for Herpetosiphon aurantiacus. Although the origin and reason of the conserved chlorimuronethyl resistance gene in field isolates of blast fungus is unclear, the ecological function of the gene is noteworthy.